Tetrahydroisoquinoline and β-Carboline Alkaloids from Haloxylon articulatum

(Cav.) Bunge (Chenopodiaceae)

Assem El-Shazlyb and Michael Winka,*
a
Institut für Pharmazie und Molekulare Biotechnologie, Im Neuenheimer Feld 364,
69120 Heidelberg, Germany. E-mail: wink@uni-hd.de
b
Department of Pharmacognosy, Faculty of Pharmacy, Zagazig University, Zagazig 44519,
Egypt
* Author for correspondence and reprint requests
Z. Naturforsch. 58 c, 477Ð480 (2003); received March 5/March 25, 2003
Salsolidine and 2-methyl,1,2,3,4-tetrahydro-β-carboline were isolated from Haloxylon arti-
culatum (growing in Egypt) besides the known carnegine and N-methylisosalsoline. 2-
methyl,1,2,3,4-tetrahydro-β-carboline was recorded for the first time in the genus Haloxylon.
The alkaloids were identified by spectroscopic methods (MS, 1D and 2D NMR).
Key words: Haloxylon articulatum, Chenopodiaceae, Tetrahydroisoquinoline Alkaloids

Introduction We have investigated the alkaloidal constituents

Haloxylon articulatum (Cav.) Bunge (family
Chenopodiaceae) is a glabrous, grey-brown,
woody, dwarf shrub usually turning darker or
blackish when dried. The plant grows wild in dry
habitats of the Mediterranean region and the Near
East. Carnegine and N-methylisosalsoline have
been previously isolated from this plant (Carling
and Sandberg, 1970). Haloxylon (Hammada) arti-
culatum ssp. scoparium from Algeria has been re-
ported to contain the alkaloids carnegine and N-
methylisosalsoline as major tetrahydroisoquino-
line alkaloids in addition to isosalsoline, salsoli-
dine, dehydrosalsolidine, isosalsolidine, N-methyl-
corydaldine, tryptamine and N-methyltryptamine
as minor alkaloids (Benkrief et al., 1990). Simple
tetrahydroisoquinoline alkaloids are common in
the Chenopodiaceae (Shamma, 1972). Further-
more, simple tetrahydroisoquinoline alkaloids can
be formed in humans and animals after alcohol
consumption since acetaldehyde (derived from
ethanol) can react with corresponding biogenic
amines. The tetrahydroisoquinoline and β-carbo-
line alkaloids affect the vegetative nervous system
(Schütte and Liebisch, 1985; Vetulani et al., 2001;
Wink et al., 1998). Simple isoquinoline and β-car-
boline alkaloids display potent, and often selective
cytotoxicity or exhibit potential antimicrobial, an-
timalarial, antiviral and anti-HIV activities (Baker,
1996; Iwasa et al., 2001).
of this desert plant from Egypt and describe the
isolation and structural determination of three
tetrahydroisoquinoline alkaloids (carnegine, salso-
lidine, 1-methylcorypalline) and for the first time
in genus Haloxylon a β-carboline alkaloid: 2-me-
thyl-1,2,3,4-tetrahydro-β-carboline.
Materials and Methods
Plant material
Aerial plant parts of Haloxylon articulatum
(Cav.) Bunge [= Hammada scoparia (Pomel) Iljin.,
Arthrophytum scoparium (Pomel) Iljin., Salsola
articulata Cav., Haloxylon scoparium Pomel] (Zo-
hary, 1966; Täckholm, 1974; Boulos, 1999) were
collected from the vicinity of El-Aresh, North
Sinia, Egypt in March 2000. The plant was kindly
identified by Dr. H. Abdel Baset, Faculty of Sci-
ence, Zagazig University. A voucher specimen was
deposited at the Herbarium of the Department of
Pharmacognosy, Faculty of Pharmacy, Zagazig
University.
Extraction and isolation of the alkaloids
About 500 g of the dried plant material were
ground to a fine powder and extracted with 80 %
EtOH in a percolator. The EtOH extract was con-
centrated in vacuo and acidified with HCl (pH 3)
and then defatted by extraction with CH2Cl2. The

0939Ð5075/2003/0700Ð0477 $ 06.00 ” 2003 Verlag der Zeitschrift für Naturforschung, Tübingen · www.znaturforsch.com · D
478 A. El-Shazly and M. Wink · Alkaloids from Haloxylon articulatum
defatted mother liquor was made alkaline with an
NH4OH solution (pH 10) and immediately ex-
tracted with CH2Cl2 to exhaustion. The latter
CH2Cl2 extract was concentrated to yield 20.1 g of
reddish-brown residue (total alkaloid). A sample
of this residue (5 g) was fractionated on a column
of neutral Al2O3 (100 g) which was packed in
CH2Cl2 and eluted with CH2Cl2- 2 % NH4OH
(25 %) in MeOH mixture adopting the gradient
elution technique to afford four alkaloids. TLC
[silica gel F254, CHCl3 Ð MeOH Ð NH4OH
(25 %), 9:1:0.5)] for the isolated alkaloids (1Ð4)
showed Rf, 0.7, 0.5, 0.48 and 0.34, respectively.
1
H- and 13C-NMR spectra were performed on
an Ac Bruker Instrument (in CDCl3 for com-
pounds 1Ð3 and CD3OD for compound 4) at 300
and 75 MHz, respectively (for data see Tables I
and II). EIMS was recorded at 70 eV by direct in-
let in JEOL (Japan). Melting points were recorded
by a Gallenkamp digital apparatus (Belton Park,
Loughborough, UK) and were uncorrected.
Carnegine (1), yellowish-white oil (2.1 g), UV
λmax (MeOH) nm 289, 354. EIMS, m/z (rel. int.):
[M+] 221 (30), 220 (34), 207 (80), 206 (100), 191
(60), 190 (76), 178 (40), 162 (50), 148 (30), 103
(35), 91 (20), 42 (14).
N-Methylisosalsoline (1-Methylcorypalline) (2),
white rosette crystals (MeOH) (20 mg), mp 170Ð
171 ∞C, UV λmax (MeOH) nm 235, 283. EIMS, m/z
(rel. int.): [M+] 207 (15), 193 (30), 192 (100), 177
(45), 164 (10), 149 (15), 121 (5), 96 (6), 91 (5), 77
(5), 57 (5), 42 (4).
Salsolidine (3), white needle-shaped crystals
(CHCI3-MeOH) (19 mg), mp 198Ð200 ∞C, UV
λmax (MeOH) nm 236, 281. EIMS, m/z (rel. int.):
[M+] 207 (78), 206 (79), 193 (80), 192 (100), 190
(60), 178 (55), 177 (60), 176 (75), 163 (30), 160
(25), 134 (23), 131 (25), 118 (20), 96 (24), 91 (25),
77 (20). 65 (10), 36 (17).
2-Methyl-1,2,3,4-tetrahydro-β-carboline (4), white
shiny needle crystals (MeOH) (50 mg), mp 213 ∞C,
UV λmax (MeOH) nm 290, 294. EIMS, m/z (rel.
int.): [M+] 186 (26), 171 (6), 144 (100), 143 (63),
115 (10), 94 (5), 77 (4), 42 (4).
Results and Discussion
Alkaloid 1 was obtained as a yellowish-white oil.
The structure of 1 was established through a com-
H3CO 4a H3CO
5 4
6 3
7 2N N
8 1 HO CH3
H3CO 8a CH3
CH3 H CH3
H
1 2
H3CO
5 4b 4a 4
6 3
N 7 2N
H3CO H 8
8 a N 8b 1 CH3
H CH3
H
4
3
Fig. 1. Structures of alkaloids found in Haloxylon articu-
latum: Carnegine (1), N-methylisosalsoline (1-methyl-
corypalline) (2), salsolidine (3), and 2-methyl-1,2,3,4-
tetrahydro-β-carboline (4).
bination of NMR and MS analysis. The EIMS
showed an [M]+ ion at 221 (for C13H19NO2) to-
gether with fragment ion peaks at m/z 206 (100 %)
[MÐMe]+ and 190 (76 %) [MÐOMe]+. The 1H-
NMR spectral data show signals for two aromatic
para-oriented protons (δ 6.51 and 6.54) and four
methyl peaks; one doublet at δ 1.32 and three sin-
glet of which one at δ 2.42 and the other two at
δ 3.79 (integrated for 6 protons). In addition to
four protons (two methylene) at δ 2.58, 2.95, 2.73
(2H) and one proton resonated at δ 3.48 coupled
with methyl group at δ 1.32. The 13C-NMR
spectrum of 1 showed 13 signals. The type of these
carbons was identified from the chemical shift and
attached proton test (APT) spectrum; four methyl
peaks, two methylene peaks, three methine peaks
and four quaternary carbons peaks. The chemical
shifts and multiplicities indicated that the com-
pound has a benzene ring with four substituents
and two methyl carbons adjacent to oxygen (δ 55.6
and 55.8), one methine (δ 58.5) and one methylene
resonated at δ 27.4. Also, there are one methyl
and one methylene adjacent to nitrogen with
δ 42.8 and 48.8, respectively. From the above men-
tioned data it was assumed that compound 1 is a
tetrahydroisoquinoline (Hughes et al., 1976; 1981).
Comparing the MS, 1H- and 13C-NMR data of this
compound with the published data (Carling and
Sandberg 1970; Khalil et al., 1992), we concluded
that this compound is carnegine. Noteworthy, car-
negine has so far previously been isolated from
a Cactus species (Brown et al., 1972; Menachery
et al., 1986).
A. El-Shazly and M. Wink · Alkaloids from Haloxylon articulatum 479

Table I. 1H-NMR chemical shifts of 1Ð4 alkaloids.*

H 1 2 3 4

1 3.48, 1H, q, J = 6.54 Hz 3.48, 1H, q, J = 6.54 Hz 4.52, 1H, q, J = 6.75 Hz 2.84, 2H, s.
3α 2.95, 1H, ddd, J = 11.7, 7.1, 2.99, 1H, ddd, J = 11.7, 7.1, 3.48, 1H, ddd, J = 11.7, 7.1,
5 Hz. 5 Hz. 5 Hz.
3β 2.58, 1H, ddd, J = 11.7, 7.1, 2.63, 1H, ddd, J = 11.7, 7.1, 3.32, 1H, ddd, J = 11.7, 7.1, {3.66, 2H, m.
5 Hz. 5 Hz. 5 Hz.
4 2.73, 2H, m. 2.76, 2H, m. 3.09, 2H, m. 2.84, 2H, m.
5 6.54, 1H, s. 6.53, 1H, s. 6.58, 1H, s. 7.37, 1H,d, J = 7.59
6 Ð Ð Ð 6.96, 1H, m.
7 Ð Ð Ð 7.03, 1H, m.
8 6.51, 1H, s. 6.65, 1H, s. 6.56, 1H, s. 7.25, 1H, d, J = 7.95
1-CH3 1.32, 3H, d, J = 6.57 Hz. 1.34, 3H, d, J = 6.57 Hz. 1.78, 3H, d, J = 6.57 Hz. Ð
N-CH3 2.42, 3H, s. 2.45, 3H, s. Ð 2.51, 3H, s.
6-OCH3 3.79, 3H, s. 3.84, 3H, s. 3.84, 3H, s. Ð
7-OCH3 3.79, 3H, s. 3.84, 3H, s. Ð

* Spectra of 1Ð3 were performed in CDCl3 and 4 was performed in CD3OD.

Table II. 13
C-NMR chemical shifts of 1Ð4 alkaloids.* loid 2 with the reported data (Khalil et al., 1992) ,
it is evident that 2 is N-methylisosalsoline (1-
C 1 2 3 4
methylcorypalline). The results showed that 1 and
1 58.5 d 58.6 d 50.7 d 54.1 t 2 were present confirming an earlier report (Car-
3 48.7 t 49.2 t 38.9 t 53.2 t ling and Sandberg, 1970) of their occurrence in
4 27.4 t 27.7 t 25.3 t 22.2 t
4a 125.8 s 125.2 s 123.4 s 107.6 s
Egyptian H. articulatum species.
4b Ð Ð Ð 128.2 s Alkaloid 3 was obtained as white needle-shaped
5 109.7 d 110.5 d 108.7 d 118.5 d crystals and its mass spectrum showed a [M]+ at
6 147.2 s 145.0 s 148.4 s 119.7 d m/z 207 (14 mass units less than in 1), correspond-
7 147.0 s 143.8 s 148.8 d 122.0 d
8 111.0 d 112.7 d 111.4 d 111.8 d ing to the formula C12H17NO2. The comparison of
1
8a 131.5 s 132.4 s 125.1 s 138.0 s H- and 13C-NMR spectrum with that of 1 (car-
8b Ð Ð Ð 132.3 s negine) revealed that 3 is N-demethylated carne-
1-CH3 19.6 q 19.7 q 20.2 q Ð
N-CH3 42.8 q 42.9 q Ð 45.6 q gine (1-methyl, 6,7-dimethoxy-1,2,3,4-tetrahydro-
6-OCH3 55.8 q 55.9 q 56.1 q Ð isoquinoline). The MS, 1H- and 13C-NMR data
7-OCH3 55.6 q Ð 55.9 q Ð (Menachery et al., 1986; Khalil, 1994) of 3 were
* Spectra of 1Ð3 were performed in CDCl3 and 4 was identical with that reported for salsolidine.
performed in CD3OD. The structure of 4 was determined from the
EIMS (which revealed a [M]+ at m/z 186, consis-
tence with C12H14N2) and a series of 1D and 2D
Alkaloid 2 had white needle-shaped crystals NMR spectroscopic experiments. The 1H-NMR
with 14 mass units less than in 1 and showed a spectrum of 4 contained signals for 14 non-ex-
fragmentation pattern (M+, 207) and a base peak changeable protons, of which four were aromatic
of m/z 192 (M+ Ð 15). In order to corroborate the while the other ten were 3 methylene (6 H), N-
structure of 2, its 1H- and 13C-NMR spectra were methyl and a nitrogen-adjacent proton. The 13H-
recorded and compared with those of structurally NMR spectrum of 4 contained twelve carbon reso-
related compounds, such as carnegine 1. 1H- and nances, of which eight arose from protonated car-
13 bons. The chemical shifts together with the
C-NMR general features were similar to those
of 1, except for the absence of one methyl peak HETCOR experiments suggested a β-carboline nu-
adjacent to oxygen. Therefore, alkaloid 2 must cleus with a 1,2,3,4-tetrahydro substitution (Prinsep
have one methoxy group and one free hydroxyl et al., 1991). The MS and 13C-NMR were identical
group instead of two methoxy groups in 1. Com- to those reported in the literature (Gander et al.,
paring the MS, 1H- and 13C-NMR spectra of alka- 1976; Poupat et al., 1976) for 2-methyl-1,2,3,4-tetra-
480 A. El-Shazly and M. Wink · Alkaloids from Haloxylon articulatum
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presence of this alkaloidal class in genus Haloxy-
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